Cystathionine γ-lyase of Saccharomyces cerevisiae: Structural gene and cystathionine γ-synthase activity

B. I. Ono, N. Ishii, K. Naito, Shin-ichi Miyoshi, S. Shinoda, S. Yamamoto, S. Ohmori

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Purification of Saccharomyces cerevisiae cystathionine γ-lyase (γ-CTLase) was hampered by the presence of a protein migrating very close to it in various types of column chromatography. The enzyme and the contaminant were nevertheless separated by polyacrylamide gel electrophoresis. N-terminal amino acid sequence analysis indicated that they are coded for by CYS3(CYI1) and MET17(MET25), respectively, leading to the conclusion that CYS3 is the structural gene for γ-CTLase and that the contaminant is O-acetylserine/O-acetylhomoserine sulfhydrylase (OAS/OAH SHLase). Based on these findings, we purified γ-CTLase by the following strategy: (1) extraction of OAS/OAH SHLase from a CYS3-disrupted strain; (2) preparation of antiserum against it; (3) identification of a strain devoid of the OAS/OAH SHLase protein using this antiserum; and (4) extraction of γ-CTLase from this strain. Purified γ-CTLase had cystathionine γ-synthase (γ-CTSase) activity if O-succinylhomoserine, but not O-acetylhomoserine, was used as substrate. From this notion we discuss the evolutional relationship between S. cerevisiae γ-CTLase and Escherichia coli γ-CTSase.

Original languageEnglish
Pages (from-to)389-397
Number of pages9
JournalYeast
Volume9
Issue number4
Publication statusPublished - 1993

Fingerprint

Cystathionine
cystathionine
Lyases
lyases
structural genes
Yeast
Saccharomyces cerevisiae
Genes
antiserum
Immune Sera
Impurities
Proteins
Column chromatography
Protein Sequence Analysis
Polyacrylates
Electrophoresis
Escherichia coli
Purification
polyacrylamide gel electrophoresis
Chromatography

Keywords

  • Amino acid sequence
  • Cystathionine γ-lyase
  • OAS/OAH sulfhydrylase
  • Purification
  • Saccharomyces cerevisiae

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry
  • Biotechnology
  • Bioengineering
  • Microbiology

Cite this

Ono, B. I., Ishii, N., Naito, K., Miyoshi, S., Shinoda, S., Yamamoto, S., & Ohmori, S. (1993). Cystathionine γ-lyase of Saccharomyces cerevisiae: Structural gene and cystathionine γ-synthase activity. Yeast, 9(4), 389-397.

Cystathionine γ-lyase of Saccharomyces cerevisiae : Structural gene and cystathionine γ-synthase activity. / Ono, B. I.; Ishii, N.; Naito, K.; Miyoshi, Shin-ichi; Shinoda, S.; Yamamoto, S.; Ohmori, S.

In: Yeast, Vol. 9, No. 4, 1993, p. 389-397.

Research output: Contribution to journalArticle

Ono, BI, Ishii, N, Naito, K, Miyoshi, S, Shinoda, S, Yamamoto, S & Ohmori, S 1993, 'Cystathionine γ-lyase of Saccharomyces cerevisiae: Structural gene and cystathionine γ-synthase activity', Yeast, vol. 9, no. 4, pp. 389-397.
Ono, B. I. ; Ishii, N. ; Naito, K. ; Miyoshi, Shin-ichi ; Shinoda, S. ; Yamamoto, S. ; Ohmori, S. / Cystathionine γ-lyase of Saccharomyces cerevisiae : Structural gene and cystathionine γ-synthase activity. In: Yeast. 1993 ; Vol. 9, No. 4. pp. 389-397.
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